LOCUS       sequence-245910-        5476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad host range plasmid derived from pBBR1-MCS5 with pemI/pemK 
            toxin-antitoxin module from Xylella fastidiosa to improve stability 
            in the absence of antibiotic selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5476)
  AUTHORS   Burbank LP, Stenger DC
  TITLE     Plasmid Vectors for Xylella fastidiosa Utilizing a Toxin-Antitoxin 
            System for Stability in the Absence of Antibiotic Selection.
  JOURNAL   Phytopathology. 2016 Aug;106(8):928-36. doi: 
            10.1094/PHYTO-02-16-0097-R. Epub 2016 Jun 7.
  PUBMED    27088393
REFERENCE   2  (bases 1 to 5476)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1094/PHYTO-02-16-0097-R"; journalName: "Phytopathology"; date: 
            "2016-08"; volume: "106"; issue: "8"; pages: "928-36"
COMMENT     Sequence Label: pBBR5pemIK
FEATURES             Location/Qualifiers
     source          1..5476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             167..829
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     primer_bind     1523..1545
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1537..1554
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1538..1554
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1564..1583
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1564..1582
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1615..1631
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1615..1631
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2370..2386)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2371..2387)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2416..2436)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2416..2434)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2455..2471)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2455..2471)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2468..2490)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2479..2495
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2503..2509,2510..2527,2528..2533))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2548..2569
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2839..2844,2845..2861,2862..2867)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3056..3589
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      join(4871..5476,1..166)
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 gctcattgcg gcaccccccg caatagctca ttgcgtaggt taaagaaaat ctgtaattga
       61 ctgccacttt tacgcaacgc ataattgttg tcgcgctgcc gaaaagttgc agctgattgc
      121 gcatggtgcc gcaaccgtgc ggcaccccta ccgcatggag ataagcatgg ccacgcagtc
      181 cagagaaatc ggcattcaag ccaagaacaa gcccggtcac tgggtgcaaa cggaacgcaa
      241 agcgcatgag gcgtgggccg ggcttattgc gaggaaaccc acggcggcaa tgctgctgca
      301 tcacctcgtg gcgcagatgg gccaccagaa cgccgtggtg gtcagccaga agacactttc
      361 caagctcatc ggacgttctt tgcggacggt ccaatacgca gtcaaggact tggtggccga
      421 gcgctggatc tccgtcgtga agctcaacgg ccccggcacc gtgtcggcct acgtggtcaa
      481 tgaccgcgtg gcgtggggcc agccccgcga ccagttgcgc ctgtcggtgt tcagtgccgc
      541 cgtggtggtt gatcacgacg accaggacga atcgctgttg gggcatggcg acctgcgccg
      601 catcccgacc ctgtatccgg gcgagcagca actaccgacc ggccccggcg aggagccgcc
      661 cagccagccc ggcattccgg gcatggaacc agacctgcca gccttgaccg aaacggagga
      721 atgggaacgg cgcgggcagc agcgcctgcc gatgcccgat gagccgtgtt ttctggacga
      781 tggcgagccg ttggagccgc cgacacgggt cacgctgccg cgccggtagc acttgggttg
      841 cgcagcaacc cgtaagtgcg ctgttccaga ctatcggctg tagccgcctc gccgccctat
      901 accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa
      961 tggaagccgg cggcacctcg ctaacggatt caccgttttt atcaggctct gggaggcaga
     1021 ataaatgatc atatcgtcaa ttattacctc cacggggaga gcctgagcaa actggcctca
     1081 ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca
     1141 atagacataa gcggctattt aacgaccctg ccctgaaccg acgaccgggt cgaatttgct
     1201 ttcgaatttc tgccattcat ccgcttatta tcacttattc aggcgtagca accaggcgtt
     1261 taagggcacc aataactgcc ttaaaaaaat tacgccccgc cctgccactc atcgcagtac
     1321 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     1381 attaacgctt acaatttcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     1441 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     1501 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc
     1561 gcgtaatacg actcactata gggcgaattg gagctccacc gcggtggcgg ccgctctaga
     1621 actagtggat cccccgggct gcaggaattc gattttcgct ataagcgtat aaatggagga
     1681 attatattat actttaatat aacttaacga aagaagccca tcatgcacac cacaaacctt
     1741 cgtaaagtcg gtggatcgat catgctggcc gttccccccg ctttccttga tcagcttcat
     1801 cttgaggtcg gcgcaacggt cggcttagcc gtcacagacg gtcgtctggt cattgagcca
     1861 gtcctacatc ctcagtacac gctggatcaa ctgctggccg aagcggaagc ctccggggct
     1921 tacccgttgc cgcccgagga acgtgaatgg gtcgatgctc cagcagtagg acgcgaactg
     1981 atatgaaacg cggcgatgtt tacatggtcg atctggagcc aaccgccgga cacgaacaac
     2041 gcggtcaccg tccggtggtt gttatctcca gtgagcgatt caatcggctt acctcttgtc
     2101 cggtgatcct tccaatcaca aacggcggcg agttcgcctc ccgcctcggg ttcgctgtgg
     2161 aattactagg tacgattaca acgggcgtcg ttcgatgtga tcaaccacgt gcgctcgact
     2221 tactcgctcg caatgcccgc aaagtggaaa gcctaccacc aaacatcttg gccgatgtct
     2281 tggcaaaagc tgtcaccatc ttccagtaat ttttggggca agttcaggcc actgcaacca
     2341 atcactagtg aattcgatat caagcttatc gataccgtcg acctcgaggg ggggcccggt
     2401 acccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     2461 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2521 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2581 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2641 acgcgcgggg agaggcggtt tgcgtattgg gcgcatgcat aaaaactgtt gtaattcatt
     2701 aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg
     2761 catcagcacc ttgtcgcctt gcgtataata tttgcccatg gacgcacacc gtggaaacgg
     2821 atgaaggcac gaacccagtt gacataagcc tgttcggttc gtaaactgta atgcaagtag
     2881 cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca
     2941 gtggcggttt tcatggcttg ttatgactgt ttttttgtac agtctatgcc tcgggcatcc
     3001 aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt
     3061 acgcagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taggtggctc
     3121 aagtatgggc atcattcgca catgtaggct cggccctgac caagtcaaat ccatgcgggc
     3181 tgctcttgat cttttcggtc gtgagttcgg agacgtagcc acctactccc aacatcagcc
     3241 ggactccgat tacctcggga acttgctccg tagtaagaca ttcatcgcgc ttgctgcctt
     3301 cgaccaagaa gcggttgttg gcgctctcgc ggcttacgtt ctgcccaagt ttgagcagcc
     3361 gcgtagtgag atctatatct atgatctcgc agtctccgga gagcaccgga ggcagggcat
     3421 tgccaccgcg ctcatcaatc tcctcaagca tgaggccaac gcgcttggtg cttatgtgat
     3481 ctacgtgcaa gcagattacg gtgacgatcc cgcagtggct ctctatacaa agttgggcat
     3541 acgggaagaa gtgatgcact ttgatatcga cccaagtacc gccacctaac aattcgttca
     3601 agccgagatc ggcttcccgg ccgcggagtt gttcggtaaa ttgtcacaac gccgcaggtg
     3661 gcacttttcg gggaaatgtg cgcgcccgcg ttcctgctgg cgctgggcct gtttctggcg
     3721 ctggacttcc cgctgttccg tcagcagctt ttcgcccacg gccttgatga tcgcggcggc
     3781 cttggcctgc atatcccgat tcaacggccc cagggcgtcc agaacgggct tcaggcgctc
     3841 ccgaaggtct cgggccgtct cttgggcttg atcggccttc ttgcgcatct cacgcgctcc
     3901 tgcggcggcc tgtagggcag gctcataccc ctgccgaacc gcttttgtca gccggtcggc
     3961 cacggcttcc ggcgtctcaa cgcgctttga gattcccagc ttttcggcca atccctgcgg
     4021 tgcataggcg cgtggctcga ccgcttgcgg gctgatggtg acgtggccca ctggtggccg
     4081 ctccagggcc tcgtagaacg cctgaatgcg cgtgtgacgt gccttgctgc cctcgatgcc
     4141 ccgttgcagc cctagatcgg ccacagcggc cgcaaacgtg gtctggtcgc gggtcatctg
     4201 cgctttgttg ccgatgaact ccttggccga cagcctgccg tcctgcgtca gcggcaccac
     4261 gaacgcggtc atgtgcgggc tggtttcgtc acggtggatg ctggccgtca cgatgcgatc
     4321 cgccccgtac ttgtccgcca gccacttgtg cgccttctcg aagaacgccg cctgctgttc
     4381 ttggctggcc gacttccacc attccgggct ggccgtcatg acgtactcga ccgccaacac
     4441 agcgtccttg cgccgcttct ctggcagcaa ctcgcgcagt cggcccatcg cttcatcggt
     4501 gctgctggcc gcccagtgct cgttctctgg cgtcctgctg gcgtcagcgt tgggcgtctc
     4561 gcgctcgcgg taggcgtgct tgagactggc cgccacgttg cccattttcg ccagcttctt
     4621 gcatcgcatg atcgcgtatg ccgccatgcc tgcccctccc ttttggtgtc caaccggctc
     4681 gacgggggca gcgcaaggcg gtgcctccgg cgggccactc aatgcttgag tatactcact
     4741 agactttgct tcgcaaagtc gtgaccgcct acggcggctg cggcgcccta cgggcttgct
     4801 ctccgggctt cgccctgcgc ggtcgctgcg ctcccttgcc agcccgtgga tatgtggacg
     4861 atggccgcga gcggccaccg gctggctcgc ttcgctcggc ccgtggacaa ccctgctgga
     4921 caagctgatg gacaggctgc gcctgcccac gagcttgacc acagggattg cccaccggct
     4981 acccagcctt cgaccacata cccaccggct ccaactgcgc ggcctgcggc cttgccccat
     5041 caattttttt aattttctct ggggaaaagc ctccggcctg cggcctgcgc gcttcgcttg
     5101 ccggttggac accaagtgga aggcgggtca aggctcgcgc agcgaccgcg cagcggcttg
     5161 gccttgacgc gcctggaacg acccaagcct atgcgagtgg gggcagtcga agggcgaagc
     5221 ccgcccgcct gccccccgag cctcacggcg gcgagtgcgg gggttccaag ggggcagcgc
     5281 caccttgggc aaggccgaag gccgcgcagt cgatcaacaa gccccggagg ggccactttt
     5341 tgccggaggg ggagccgcgc cgaaggcgtg ggggaacccc gcaggggtgc ccttctttgg
     5401 gcaccaaaga actagatata gggcgaaatg cgaaagactt aaaaatcaac aacttaaaaa
     5461 aggggggtac gcaaca
//
